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. 1986 Mar 11;14(5):2391–2405. doi: 10.1093/nar/14.5.2391

Polymerase-defective mutant of the Bryan high-titer strain of Rous sarcoma virus.

M Sudol, T L Lerner, H Hanafusa
PMCID: PMC339665  PMID: 2421248

Abstract

A mutant of the Bryan high-titer strain of Rous sarcoma virus defective in reverse transcriptase is known as type alpha (BH-RSV alpha). BH-RSV alpha virion particles do not contain any polymerase-related proteins but they direct the synthesis of a normal sized Pr180 gag-pol polyprotein precursor in infected cells. Using a bioassay for polymerase gene function that is based on the requirement of viral replication for transformation of transfected chicken cells, we have localized the defect to the 2.5 kb EcoRI-KpnI DNA fragment containing more than 90% of the polymerase gene by comparison with the corresponding DNA fragment from the wild-type polymerase-positive BH-RSV, called type beta. In vitro recombination experiments with the polymerase gene of Schmidt-Ruppin RSV allowed us to map the defect to the 0.86 kb XbaI-BglII DNA fragment of the BH-RSV alpha polymerase. DNA sequence analysis of the entire polymerase gene of BH-RSV alpha and beta has revealed one point mutation that maps within that XbaI-BglII fragment and substitutes leucine in BH-RSV alpha for glutamine in the wild-type BH-RSV beta.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BRYAN W. R., CALNAN D., MOLONEY J. B. Biological studies on the Rous sarcoma virus. III. The recovery of virus from experimental tumors in relation to initiating dose. J Natl Cancer Inst. 1955 Aug;16(1):317–335. [PubMed] [Google Scholar]
  2. BRYAN W. R. Enhancement of virus yield of Rous sarcomas. Acta Unio Int Contra Cancrum. 1959;15:764–767. [PubMed] [Google Scholar]
  3. Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
  4. Cross F. R., Hanafusa H. Local mutagenesis of Rous sarcoma virus: the major sites of tyrosine and serine phosphorylation of pp60src are dispensable for transformation. Cell. 1983 Sep;34(2):597–607. doi: 10.1016/0092-8674(83)90392-6. [DOI] [PubMed] [Google Scholar]
  5. Duesberg P. H., Kawai S., Wang L. H., Vogt P. K., Murphy H. M., Hanafusa H. RNA of replication-defective strains of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1569–1573. doi: 10.1073/pnas.72.4.1569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HANAFUSA H., HANAFUSA T., RUBIN H. ANALYSIS OF THE DEFECTIVENESS OF ROUS SARCOMA VIRUS, II. SPECIFICATION OF RSV ANTIGENICITY BY HELPER VIRUS. Proc Natl Acad Sci U S A. 1964 Jan;51:41–48. doi: 10.1073/pnas.51.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HANAFUSA H., HANAFUSA T., RUBIN H. The defectiveness of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1963 Apr;49:572–580. doi: 10.1073/pnas.49.4.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hanafusa H., Baltimore D., Smoler D., Watson K. F., Yaniv A., Spiegelman S. Absence of polymerase protein in virions of alpha-type rous sarcoma virus. Science. 1972 Sep 29;177(4055):1188–1191. doi: 10.1126/science.177.4055.1188. [DOI] [PubMed] [Google Scholar]
  9. Hanafusa H., Hanafusa T. Further studies on RSV production form transformed cells. Virology. 1968 Apr;34(4):630–636. doi: 10.1016/0042-6822(68)90084-6. [DOI] [PubMed] [Google Scholar]
  10. Hanafusa H., Hanafusa T. Noninfectious RSV deficient in DNA polymerase. Virology. 1971 Jan;43(1):313–316. doi: 10.1016/0042-6822(71)90251-0. [DOI] [PubMed] [Google Scholar]
  11. Hanafusa H., Miyamoto T., Hanafusa T. A cell-associated factor essential for formation of an infectious form of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1970 Jun;66(2):314–321. doi: 10.1073/pnas.66.2.314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hanafusa T., Miyamoto T., Hanafusa H. A type of chick embryo cell that fails to support formation of infectious RSV. Virology. 1970 Jan;40(1):55–64. doi: 10.1016/0042-6822(70)90378-8. [DOI] [PubMed] [Google Scholar]
  13. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lerner T. L., Hanafusa H. DNA sequence of the Bryan high-titer strain of Rous sarcoma virus: extent of env deletion and possible genealogical relationship with other viral strains. J Virol. 1984 Feb;49(2):549–556. doi: 10.1128/jvi.49.2.549-556.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  17. Oppermann H., Bishop J. M., Varmus H. E., Levintow L. A joint produce of the genes gag and pol of avian sarcoma virus: a possible precursor of reverse transcriptase. Cell. 1977 Dec;12(4):993–1005. doi: 10.1016/0092-8674(77)90164-7. [DOI] [PubMed] [Google Scholar]
  18. Panet A., Baltimore D., Hanafusa T. Quantitation of avian RNA tumor virus reverse transcriptase by radioimmunoassay. J Virol. 1975 Jul;16(1):146–152. doi: 10.1128/jvi.16.1.146-152.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sawyer R. C., Hanafusa H. Comparison of the small RNAs of polymerase-deficient and polymerase-positive Rous sarcoma virus and another species of avian retrovirus. J Virol. 1979 Mar;29(3):863–871. doi: 10.1128/jvi.29.3.863-871.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scheele C. M., Hanafusa H. Proteins of helper-dependent RSV. Virology. 1971 Aug;45(2):401–410. doi: 10.1016/0042-6822(71)90341-2. [DOI] [PubMed] [Google Scholar]
  21. Schwartz D. E., Tizard R., Gilbert W. Nucleotide sequence of Rous sarcoma virus. Cell. 1983 Mar;32(3):853–869. doi: 10.1016/0092-8674(83)90071-5. [DOI] [PubMed] [Google Scholar]
  22. Staden R. Sequence data handling by computer. Nucleic Acids Res. 1977 Nov;4(11):4037–4051. doi: 10.1093/nar/4.11.4037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Witte O. N., Baltimore D. Relationship of retrovirus polyprotein cleavages to virion maturation studied with temperature-sensitive murine leukemia virus mutants. J Virol. 1978 Jun;26(3):750–761. doi: 10.1128/jvi.26.3.750-761.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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